RNA Detection

2.8 Microscopy 1. Inverted confocal microscope with a low and a high magnifica-
tion, high numerical aperture objective (e.g., Leica SP8 with a
20 dry and a 631.4 NA objective).

2. 488 nm or 514 nm laser lines to excite thiazole orange (TO).


3. 561 nm or 594 nm laser lines to excite quinoline blue (QB).

Fig. 1Preparation of the Drosophila egg chambers for in vivo injection and

imaging. (a) Separation and alignment of the ovarioles at the cultivating med-

ium–halocarbon oil interface with two tungsten needles. After the separation is

finished, the excess of medium is blotted away. (b) Schematics of the incised

plastic slide. The coverslip is held by a thin plastic sheet cut in a U-shape (thin

outline) that is secured to the plastic slide (thick outline) by two strips of dual

faced adhesive tapes (gray rectangles)

In vivo mRNP Detection by FIT Probes 277